苏云金芽胞杆菌Cry Ⅰ基因RNA探针载体的构建

CONSTRUCTION OF RNA PROBE VECTOR OF BACILLUS THURINGIENSIS CryⅠGENES

将苏云金芽胞杆菌杀虫晶体蛋白ＣｒｙⅠＡ基因的高保守区ＥｃｏＲＩ－Ｆ片段重组到ｐＳＥＬＥＣＴ－１载体的ＥｃｏＲＩ位点，构建出能够高效转录ＲＮＡ的探针载体ｐＳＢＰＬ－１。该载体在Ｔ７ＲＮＡ聚合酶作用下，通过ＤＩＧ体外标记转录制备的ＲＮＡ探针，具有灵敏度高，背景清楚，省时等优点，为苏云金芽胞杆菌分子生物学研究，及对鳞翅目有特异毒性高效菌株的筛选提供了一种方便而有效的方法。

High conserved domain,EcoRⅠ-F fragment,of Bacillus thuringiensis insecticidal crystal protein CryⅠA gene was ligated into EcoRI site of pSELECT-1 vector.The constructed RNA probe vector, pSBL-1, could transcribe RNA at high efficiency. The RNA probe labeled by DIG transcription system in vitro had greater yield,higher sensitivity and lower background than DNA probe. It supplies an effective method for molecular biology study of Bacillus thuringiensis and screening lepidopteran-specific strains.